Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Biochemistry. 2012 Sep 4;51(35):6968-80. doi: 10.1021/bi3003023. Epub 2012 Aug 23.
One of the most intriguing questions confronting the bone morphogenetic protein family is the mechanism of ligand recognition, because there are more ligands than receptors. Crystal structures of two type II receptors, ActR-II and BMPR-II, are essentially identical, and a loop structure (A-loop) has been suggested to play a role in determining ligand specificity. A solution biophysical study showed mutations of several A-loop residues in these two receptors exert different ligand binding effects. Thus, the issues of mechanism of ligand recognition and specificity remain unresolved. We examined effects of mutations of residues Y40, G47, and S107 in BMPR-II. These residues are not identified as being in contact with the ligand in the BMP-7-BMPR-II complex but are found mutated in genetic diseases. They are likely to be useful in identifying their roles in differentiating the various BMP ligands. Spectroscopic probing revealed little mutation-induced structural change in BMPR-II. Ligand binding studies revealed that Y40 plays a significant role in differentiating three distinct ligands; G47 and S107 affect ligand binding to a lesser extent. The role of the A-loop in ActR-II or BMPR-II is dependent on the host sequence of the receptor extracellular domain (ECD) in which it is embedded, suggesting a host-guest relationship between the A-loop and the rest of the ECD. Computational analysis demonstrated a long-range connectivity between Y40, G47, and S107 and other locations in BMPR-II. An integration of these results on functional energetics and protein structures clearly demonstrates, for the first time, an intradomain communication network within BMPR-II.
骨形态发生蛋白家族面临的最有趣的问题之一是配体识别的机制,因为配体比受体多。两种 II 型受体 ActR-II 和 BMPR-II 的晶体结构基本相同,并且已经提出了一个环结构(A 环)在决定配体特异性方面起作用。一项溶液生物物理研究表明,这两种受体中的几个 A 环残基的突变会产生不同的配体结合效应。因此,配体识别和特异性的机制问题仍然没有解决。我们检查了 BMPR-II 中残基 Y40、G47 和 S107 突变的影响。这些残基在 BMP-7-BMPR-II 复合物中未被确定为与配体接触,但在遗传疾病中发现发生了突变。它们可能有助于确定它们在区分各种 BMP 配体中的作用。光谱探测显示 BMPR-II 的突变引起的结构变化很小。配体结合研究表明,Y40 在区分三种不同的配体方面起着重要作用;G47 和 S107 对配体结合的影响较小。A 环在 ActR-II 或 BMPR-II 中的作用取决于其嵌入的受体细胞外结构域(ECD)的宿主序列,这表明 A 环与 ECD 的其余部分之间存在主客体关系。计算分析表明,Y40、G47 和 S107 与 BMPR-II 中的其他位置之间存在长程连通性。这些结果在功能能量学和蛋白质结构上的综合,首次清楚地表明了 BMPR-II 内的一个域内通讯网络。